Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

  • Letter
  • Published:

Cis-interactions between Notch and Delta generate mutually exclusive signalling states

Nature volume 465, pages 86–90 (2010)Cite this article

Subjects

Abstract

The Notch–Delta signalling pathway allows communication between neighbouring cells during development1. It has a critical role in the formation of ‘fine-grained’ patterns, generating distinct cell fates among groups of initially equivalent neighbouring cells and sharply delineating neighbouring regions in developing tissues2,3,4,5. The Delta ligand has been shown to have two activities: it transactivates Notch in neighbouring cells and cis-inhibits Notch in its own cell. However, it remains unclear how Notch integrates these two activities and how the resulting system facilitates pattern formation. Here we report the development of a quantitative time-lapse microscopy platform for analysing Notch–Delta signalling dynamics in individual mammalian cells, with the aim of addressing these issues. By controlling both cis- and trans-Delta concentrations, and monitoring the dynamics of a Notch reporter, we measured the combined cis–trans input–output relationship in the Notch–Delta system. The data revealed a striking difference between the responses of Notch to trans- and cis-Delta: whereas the response to trans-Delta is graded, the response to cis-Delta is sharp and occurs at a fixed threshold, independent of trans-Delta. We developed a simple mathematical model that shows how these behaviours emerge from the mutual inactivation of Notch and Delta proteins in the same cell. This interaction generates an ultrasensitive switch between mutually exclusive sending (high Delta/low Notch) and receiving (high Notch/low Delta) signalling states. At the multicellular level, this switch can amplify small differences between neighbouring cells even without transcription-mediated feedback. This Notch–Delta signalling switch facilitates the formation of sharp boundaries and lateral-inhibition patterns in models of development, and provides insight into previously unexplained mutant behaviours.

This is a preview of subscription content, access via your institution

Access options

Buy this article

  • Purchase on SpringerLink
  • Instant access to full article PDF
Buy now

Prices may be subject to local taxes which are calculated during checkout

Figure 1: System for analysing signal integration in the Notch–Delta pathway.
Figure 2: Transactivation of Notch occurs in a graded fashion.
Figure 3: Cis – trans signal integration by Notch.
Figure 4: The mutual-inactivation model in multicellular patterning.

Similar content being viewed by others

References

  1. Artavanis-Tsakonas, S., Rand, M. D. & Lake, R. J. Notch signaling: cell fate control and signal integration in development. Science 284, 770–776 (1999)

    Article  ADS  CAS  PubMed  Google Scholar 

  2. Goodyear, R. & Richardson, G. Pattern formation in the basilar papilla: evidence for cell rearrangement. J. Neurosci. 17, 6289–6301 (1997)

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  3. Heitzler, P. & Simpson, P. The choice of cell fate in the epidermis of Drosophila. Cell 64, 1083–1092 (1991)

    Article  CAS  PubMed  Google Scholar 

  4. Huppert, S. S., Jacobsen, T. L. & Muskavitch, M. A. Feedback regulation is central to Delta-Notch signalling required for Drosophila wing vein morphogenesis. Development 124, 3283–3291 (1997)

    CAS  PubMed  Google Scholar 

  5. de Celis, J. F., Bray, S. & Garcia-Bellido, A. Notch signalling regulates veinlet expression and establishes boundaries between veins and interveins in the Drosophila wing. Development 124, 1919–1928 (1997)

    CAS  PubMed  Google Scholar 

  6. Bray, S. J. Notch signalling: a simple pathway becomes complex. Nature Rev. Mol. Cell Biol. 7, 678–689 (2006)

    Article  CAS  Google Scholar 

  7. de Celis, J. F. & Bray, S. Feed-back mechanisms affecting Notch activation at the dorsoventral boundary in the Drosophila wing. Development 124, 3241–3251 (1997)

    CAS  PubMed  Google Scholar 

  8. Micchelli, C. A., Rulifson, E. J. & Blair, S. S. The function and regulation of cut expression on the wing margin of Drosophila: Notch, Wingless and a dominant negative role for Delta and Serrate. Development 124, 1485–1495 (1997)

    CAS  PubMed  Google Scholar 

  9. Klein, T., Brennan, K. & Arias, A. M. An intrinsic dominant negative activity of serrate that is modulated during wing development in Drosophila. Dev. Biol. 189, 123–134 (1997)

    Article  CAS  PubMed  Google Scholar 

  10. Miller, A. C., Lyons, E. L. & Herman, T. G. cis-Inhibition of notch by endogenous delta biases the outcome of lateral inhibition. Curr. Biol. 19, 1378–1383 (2009)

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  11. Cordle, J. et al. A conserved face of the Jagged/Serrate DSL domain is involved in Notch trans-activation and cis-inhibition. Nature Struct. Mol. Biol. 15, 849–857 (2008)

    Article  CAS  Google Scholar 

  12. Matsuda, M. & Chitnis, A. B. Interaction with Notch determines endocytosis of specific Delta ligands in zebrafish neural tissue. Development 136, 197–206 (2009)

    Article  CAS  PubMed  Google Scholar 

  13. Kakidani, H. & Ptashne, M. GAL4 activates gene expression in mammalian cells. Cell 52, 161–167 (1988)

    Article  CAS  PubMed  Google Scholar 

  14. Struhl, G. & Adachi, A. Nuclear access and action of notch in vivo. Cell 93, 649–660 (1998)

    Article  CAS  PubMed  Google Scholar 

  15. Aster, J. C. et al. Essential roles for ankyrin repeat and transactivation domains in induction of T-cell leukemia by notch1. Mol. Cell. Biol. 20, 7505–7515 (2000)

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  16. Yang, L. T. et al. Fringe glycosyltransferases differentially modulate Notch1 proteolysis induced by Delta1 and Jagged1. Mol. Biol. Cell 16, 927–942 (2005)

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  17. Rothenberg, E. V., Moore, J. E. & Yui, M. A. Launching the T-cell-lineage developmental programme. Nature Rev. Immunol. 8, 9–21 (2008)

    Article  CAS  Google Scholar 

  18. Varnum-Finney, B. et al. Immobilization of Notch ligand, Delta-1, is required for induction of notch signaling. J. Cell Sci. 113, 4313–4318 (2000)

    CAS  PubMed  Google Scholar 

  19. Wang, S. et al. Notch receptor activation inhibits oligodendrocyte differentiation. Neuron 21, 63–75 (1998)

    Article  PubMed  Google Scholar 

  20. Rosenfeld, N., Young, J. W., Alon, U., Swain, P. S. & Elowitz, M. B. Gene regulation at the single-cell level. Science 307, 1962–1965 (2005)

    Article  ADS  CAS  PubMed  Google Scholar 

  21. Jacobsen, T. L., Brennan, K., Arias, A. M. & Muskavitch, M. A. Cis-interactions between Delta and Notch modulate neurogenic signalling in Drosophila. Development 125, 4531–4540 (1998)

    CAS  PubMed  Google Scholar 

  22. Shaye, D. D. & Greenwald, I. LIN-12/Notch trafficking and regulation of DSL ligand activity during vulval induction in Caenorhabditis elegans. Development 132, 5081–5092 (2005)

    Article  CAS  PubMed  Google Scholar 

  23. de Celis, J. F. & Bray, S. J. The Abruptex domain of Notch regulates negative interactions between Notch, its ligands and Fringe. Development 127, 1291–1302 (2000)

    CAS  PubMed  Google Scholar 

  24. Collier, J. R., Monk, N. A., Maini, P. K. & Lewis, J. H. Pattern formation by lateral inhibition with feedback: a mathematical model of Delta-Notch intercellular signalling. J. Theor. Biol. 183, 429–446 (1996)

    Article  CAS  PubMed  Google Scholar 

  25. Plahte, E. Pattern formation in discrete cell lattices. J. Math. Biol. 43, 411–445 (2001)

    Article  MathSciNet  CAS  PubMed  Google Scholar 

  26. Melen, G. J., Levy, S., Barkai, N. & Shilo, B. Z. Threshold responses to morphogen gradients by zero-order ultrasensitivity. Mol. Syst. Biol. 1 10.1038/msb4100036 (2005)

  27. Levine, E., Zhang, Z., Kuhlman, T. & Hwa, T. Quantitative characteristics of gene regulation by small RNA. PLoS Biol. 5, e229 (2007)

    Article  PubMed Central  PubMed  Google Scholar 

  28. Buchler, N. E. & Louis, M. Molecular titration and ultrasensitivity in regulatory networks. J. Mol. Biol. 384, 1106–1119 (2008)

    Article  CAS  PubMed  Google Scholar 

  29. Lenz, D. H. et al. The small RNA chaperone Hfq and multiple small RNAs control quorum sensing in Vibrio harveyi and Vibrio cholerae. Cell 118, 69–82 (2004)

    Article  CAS  PubMed  Google Scholar 

Download references

Acknowledgements

We would like to thank I. Bernstein for the IgG-Deltaext, U. Lendahl for the 12xCSL reporter construct, J. Aster for human NOTCH1 and other constructs, and G. Weinmaster for the rat DLL1 construct and advice. We also thank R. Tsien and K. Thorn for mCherry, S. Megason and S. Fraser for H2B–citrine and other constructs, R. Diamond and D. Perez for assistance with FACS, and F. Tan and J. Yong for help with cloning some of the constructs. We thank A. Eldar, J. Locke, G. Seelig, R. Kishony, B. Shraiman, A. C. Oates and members of the Elowitz laboratory for discussions and advice. This work was supported by the US National Institutes of Health Fellowship F32GM77014 (D.S.), the Caltech Center for Biological Circuit Design and the Packard Foundation. A.L. acknowledges support from the Fannie and John Hertz Foundation and the UCLA/Caltech Medical Scientist Training Program (NIH GM08042). J.G.O. acknowledges support from the Ministerio de Ciencia e Innovacion (Spain, project FIS2009-13360 and the I3 programme).

Author information

Authors and Affiliations

  1. Division of Biology and Department of Applied Physics, Howard Hughes Medical Institute, California Institute of Technology, 1200 East California Boulevard, Pasadena, California 91125, USA,

    David Sprinzak, Amit Lakhanpal, Lauren LeBon, Leah A. Santat, Michelle E. Fontes & Michael B. Elowitz

  2. Department of Chemical and Systems Biology, Stanford University School of Medicine, 269 Campus Drive, Stanford, California 94305, USA,

    Graham A. Anderson

  3. Department de Física i Enginyeria Nuclear, Universitat Politècnica de Catalunya, Colom 11, E-08222 Terrassa, Spain,

    Jordi Garcia-Ojalvo

Authors
  1. David Sprinzak
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Amit Lakhanpal
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Lauren LeBon
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Leah A. Santat
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Michelle E. Fontes
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Graham A. Anderson
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Jordi Garcia-Ojalvo
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Michael B. Elowitz
    View author publications

    You can also search for this author in PubMed Google Scholar

Contributions

D.S. and M.B.E. designed the research. D.S., L.A.S., M.E.F. and G.A.A. built cell lines and performed experiments. D.S., A.L., L.L., J.G.-O. and M.B.E. performed data analysis and mathematical modelling. D.S. and M.B.E. wrote the manuscript with substantial input from the other authors.

Corresponding author

Correspondence to Michael B. Elowitz.

Ethics declarations

Competing interests

The authors declare no competing financial interests.

Supplementary information

Supplementary Information

This file contains Supplementary Figures S1-S17 with legends, Supplementary Tables S1-S3, Supplementary Methods and Data Analysis and Data and References for the Supplementary Material. (PDF 3625 kb)

Supplementary Movie 1

This movie shows the trans-activation of hN1G4esn by plate bound Delta. Movie used to generate filmstrip in Figure 2B. (AVI 10479 kb)

Supplementary Movie 2

This movie shows the effect of cis-Delta on hN1G4esn activation. Movie used to generate filmstrip in Figure 3B. (AVI 22000 kb)

Supplementary Movie 3

This movie shows the trans-activation of hN1G4esn-No-Delta by co-culture with Delta-expressing cells. Movie used to generate filmstrip in Fig S5. (AVI 5340 kb)

PowerPoint slides

Rights and permissions

Reprints and permissions

About this article

Cite this article

Sprinzak, D., Lakhanpal, A., LeBon, L. et al. Cis-interactions between Notch and Delta generate mutually exclusive signalling states. Nature 465, 86–90 (2010). https://doi.org/10.1038/nature08959

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/nature08959

This article is cited by

Comments

By submitting a comment you agree to abide by our Terms and Community Guidelines. If you find something abusive or that does not comply with our terms or guidelines please flag it as inappropriate.

Search

Advanced search

Quick links

Nature Briefing

Sign up for the Nature Briefing newsletter — what matters in science, free to your inbox daily.

Get the most important science stories of the day, free in your inbox. Sign up for Nature Briefing